Abstract
Summary: Starting from children and adult skeletal muscle biopsy specimens (150 mg), we devised a scaled-down technique for isolation and cell-free translation of total cellular messenger RNAs in a reticulocyte-lysate system. Moreover, starting from 200 mg of human muscle, the technique allowed us to purify a neosynthesized protein, glycogen phosphorylase b (EC 2.4.1.1), using a specific immunoaffinity microchromatography procedure.
Speculation: These results show that several neuromuscular diseases can now be investigated with the new tools of molecular biology, especially when the specific immunoabsorbent is available. If one considers the innocuity of a needle (80 mg of tissue per puncture) or surgical biopsy taking such small amounts of muscle, it is tempting to imagine the potential applications of these techniques for studying the mechanism of several inherited childhood metabolic diseases and for investigating pathologic changes in gene expression.
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Munnich, A., Daegelen, D., Besmond, C. et al. Cell-Free Translation of Messenger RNAs from Human Muscle Biopsies: a Miniaturized Tool for Investigation of Neuromuscular Diseases. Pediatr Res 16, 335–339 (1982). https://doi.org/10.1203/00006450-198205000-00001
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DOI: https://doi.org/10.1203/00006450-198205000-00001
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